Large-scale voltage imaging in behaving mice using targeted illumination.
Large-scale voltage imaging in behaving mice using targeted illumination.
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DOI：
10.1016/j.isci.2021.103263
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发表时间：
2021-11-19
期刊：
影响因子：
5.8
通讯作者：
Han X
中科院分区：
文献类型：
--
作者：
Xiao S;Lowet E;Gritton HJ;Fabris P;Wang Y;Sherman J;Mount RA;Tseng HA;Man HY;Straub C;Piatkevich KD;Boyden ES;Mertz J;Han X
遗传编码的电压指示器的最新改进使得能够在体内对动作电位和阈下跨膜电压进行光学成像。为了在大的解剖区域内同时对许多神经元进行高速电压成像，宽视野显微镜仍然是一种必不可少的工具。然而，光学切片的缺乏使得宽视场显微镜易于背景交叉污染。我们实施了一种基于数字图像处理设备的靶向照明策略，以将照明限制在感兴趣的细胞上，并在理论上和实验上用SomArchon表达神经元量化了所得到的改善。我们发现，有针对性的照明增加SomArchon信号对比度，减少光漂白，并减少背景交叉污染。通过使用高速，大面积的sCMOS相机，我们在几分钟内同时对行为小鼠中的数十个尖峰神经元进行了常规成像。因此，这里描述的有针对性的照明策略提供了一个简单的解决方案，用于在行为动物的大视场内对许多神经元进行宽场电压成像。一种易于实现的基于数字X射线的靶向照明显微镜靶向照明改善了大脑中的电压成像信号对比度靶向照明减少了光漂白并减少了大脑中的背景行为小鼠中数十个神经元同时进行的常规电压成像光学成像;神经科学;神经科学技术
Recent improvements in genetically encoded voltage indicators enabled optical imaging of action potentials and subthreshold transmembrane voltage in vivo. To perform high-speed voltage imaging of many neurons simultaneously over a large anatomical area, widefield microscopy remains an essential tool. However, the lack of optical sectioning makes widefield microscopy prone to background cross-contamination. We implemented a digital-micromirror-device-based targeted illumination strategy to restrict illumination to the cells of interest and quantified the resulting improvement both theoretically and experimentally with SomArchon expressing neurons. We found that targeted illumination increased SomArchon signal contrast, decreased photobleaching, and reduced background cross-contamination. With the use of a high-speed, large-area sCMOS camera, we routinely imaged tens of spiking neurons simultaneously over minutes in behaving mice. Thus, the targeted illumination strategy described here offers a simple solution for widefield voltage imaging of many neurons over a large field of view in behaving animals. An easily implementable digital-micromirror-based targeted illumination microscope Targeted illumination improves voltage imaging signal contrast in the brain Targeted illumination decreases photobleaching and reduces background in the brain Routine voltage imaging of tens of neurons simultaneously in behaving mice Optical imaging; Neuroscience; Techniques in neuroscience
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